High-Performance Liquid Chromatography of Peptides and Proteins : Separation, Analysis, and Conformation

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"This book consists of a series of 82 precise, easy-to-read articles by internationally renowned scientists and emphasizes the practical approach to HPLC with minimal theory, although the underlying principles for peptide and protein separations are clearly expressed. All of the major modes of microbore, ultrafast and analytical HPLC are discussed, including size-exclusion, ion-exchange, reversed-phase, hydrophobic interaction, and affinity and immunoaffinity chromatography. A section on preparative HPLC, including displacement techniques, is also presented. Problem-solving approaches to the separation of various classes of biologically active peptides and proteins are thoroughly explored, while the importance of peptide standards for monitoring column performance and for optimizing separation conditions is emphasized. Several articles focus on the choice of the correct detection method (electrochemical, UV, fluorescence), as well as the need for a proper knowledge of approaches to column and instrument maintenance and trouble-shooting. A section on predictive approaches deals with both computer simulation of peptide separations and peptide structure. The book also includes complementary techniques to HPLC, as well as other useful applications of HPLC. It enables both novice and experienced chromatographers to realize the full potential of this extremely powerful technique, in the process making an important contribution to scientific literature."--Provided by publisher. Read more... Abstract: "This book consists of a series of 82 precise, easy-to-read articles by internationally renowned scientists and emphasizes the practical approach to HPLC with minimal theory, although the underlying principles for peptide and protein separations are clearly expressed. All of the major modes of microbore, ultrafast and analytical HPLC are discussed, including size-exclusion, ion-exchange, reversed-phase, hydrophobic interaction, and affinity and immunoaffinity chromatography. A section on preparative HPLC, including displacement techniques, is also presented. Problem-solving approaches to the separation of various classes of biologically active peptides and proteins are thoroughly explored, while the importance of peptide standards for monitoring column performance and for optimizing separation conditions is emphasized. Several articles focus on the choice of the correct detection method (electrochemical, UV, fluorescence), as well as the need for a proper knowledge of approaches to column and instrument maintenance and trouble-shooting. A section on predictive approaches deals with both computer simulation of peptide separations and peptide structure. The book also includes complementary techniques to HPLC, as well as other useful applications of HPLC. It enables both novice and experienced chromatographers to realize the full potential of this extremely powerful technique, in the process making an important contribution to scientific literature."--Provided by publisher Content: Cover Title Page Copyright Page Preface Table of Contents SECTION I: PRACTICAL ASPECTS OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY Properties of Peptides/Proteins and Practical Implications Standard Chromatographic Conditions for Size-Exclusion, Ion-Exchange, Reversed-Phase and Hydrophobic Interaction Chromatography Preventive Maintenance and Troubleshooting LC Instrumentation Tools and Techniques to Extend LC Column Lifetimes Mobile Phase Preparation and Column Maintenance HPLC Guard Columns On-Line Derivatization of Silica Supports for Regeneration of Reversed-Phase Columns SECTION II: THEORETICAL ASPECTS OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHYHPLC Terminology: Practical and Theoretical High-Performance Liquid Chromatography of Peptides and Proteins: General Principles and Basic Theory High-Performance Liquid Chromatography of Peptides and Proteins: Quantitative Relationships for Isocratic and Gradient Elution of Peptides and Proteins SECTION III: SIZE-EXCLUSION CHROMATOGRAPHY Requirement for Peptide Standards to Monitor Ideal and Non-Ideal Behavior in Size- Exclusion Chromatography High-Perfonnance Size-Exclusion Chromatography of Proteins Size-Exclusion Chromatography of ProteinsSize-Exclusion High-Performance Liquid Chromatography of Integral Membrane Proteins and the Effect of Detergents on Immunological Activity A Reversed-Phase Trace Enrichment Technique for the Loading of Size-Exclusion HPLC Columns SECTION IV: ION-EXCHANGE CHROMATOGRAPHY The Use of Peptide Standards for Monitoring Ideal and Non-Ideal Behavior in Cation-Exchange Chromatography Ion-Exchange High-Performance Liquid Chromatography of Peptides Use of Ion-Exchange Cartridges for Batch Extraction in the Purification of Pituitary Peptides High-Performance Ion-Exchange Chromatography of ProteinsHigh-Performance Ion-Exchange Chromatography of Proteins: Application to the Purification of Enzymes Anion Exchange High-Performance Liquid Chromatography of Viral Proteins Sample Preparation and Application of Different HPLC Methods for the Separation of Hydrophobic Membrane Proteins Cation-Exchange High-Performance Liquid Chromatography: Group Separation of Acidic and Basic Proteins Using Volatile Solvents Separation of Protein Isoforms: An Ion-Exchange HPLC Separation of Bovine and Cardiac Troponin Complexes Assessment of Steroid Receptor Polymorphism by High-Performance Ion-Exchange ChromatographySECTION V: REVERSED-PHASE CHROMATOGRAPHY Reversed-Phase Packings for the Separation of Peptides and Proteins by Means of Gradient Elution High-Performance Liquid Chromatography Commercially Available Columns and Packings for Reversed-Phase HPLC of Peptides and Proteins Requirement for Peptide Standards to Monitor Colunm Performance and the Effect of Column Dimensions, Organic Modifiers, and Temperature in Reversed-Phase Chromatography